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PRMT5 methylating SMAD4 activates TGF-β signaling and promotes colorectal cancer metastasis

Oncogene volume 42, pages 1572–1584 (2023)Cite this article

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Abstract

Perturbations in transforming growth factor-β (TGF-β) signaling can lead to a plethora of diseases, including cancer. Mutations and posttranslational modifications (PTMs) of the partner of SMAD complexes contribute to the dysregulation of TGF-β signaling. Here, we reported a PTM of SMAD4, R361 methylation, that was critical for SMAD complexes formation and TGF-β signaling activation. Through mass spectrometric, co-immunoprecipitation (Co-IP) and immunofluorescent (IF) assays, we found that oncogene protein arginine methyltransferase 5 (PRMT5) interacted with SMAD4 under TGF-β1 treatment. Mechanically, PRMT5 triggered SMAD4 methylation at R361 and induced SMAD complexes formation and nuclear import. Furthermore, we emphasized that PRMT5 interacting and methylating SMAD4 was required for TGF-β1-induced epithelial-mesenchymal transition (EMT) and colorectal cancer (CRC) metastasis, and SMAD4 R361 mutation diminished PRMT5 and TGF-β1-induced metastasis. In addition, highly expressed PRMT5 or high level of SMAD4 R361 methylation indicated worse outcomes in clinical specimens analysis. Collectively, our study highlights the critical interaction of PRMT5 and SMAD4 and the roles of SMAD4 R361 methylation for controlling TGF-β signaling during metastasis. We provided a new insight for SMAD4 activation. And this study indicated that blocking PRMT5-SMAD4 signaling might be an effective targeting strategy in SMAD4 wild-type CRC.

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Fig. 1: PRMT5 methylates Smad4 at R361.
Fig. 2: PRMT5 promotes the formation of SMAD complexes dependent on methylation of SMAD4 at R361.
Fig. 3: The methylation of SMAD4 at R361 facilitates SMAD complexes’ nuclear import.
Fig. 4: PRMT5 promotes TGF-β-induced EMT and metastasis of CRC in vitro.
Fig. 5: Targeting the methylation of SMAD4 at R361 inhibits CRC metastasis in vivo.
Fig. 6: The level of SMAD4 R361me2s can be used as a prognostic indicator for colorectal cancer.

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Data availability

Source data of mass spectrometric analyses are provided with this paper. Other data that support the findings are available from the corresponding author upon reasonable request.

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Acknowledgements

This study was supported by the National Key Research and Development Program of China (No. 2022YFA1105303 GW) and National Natural Science Foundation of China (NSFC) (Nos. 81922053 GW, 81974432 GW, 81874186 JH, No. 82273254 JH, No. 82260521 XS) and Hubei Provincial Finance Department (SCZ202203 GW). We would like to acknowledge the support from the Molecular Medical Center of Tongji Hospital, Huazhong University of Science and Technology. We appreciate the assistance from members of Guihua Wang’s laboratory and Junbo Hu’s laboratory. We thank the Protein Chemistry and Proteomics Facility, Tsinghua University for the contribution of mass spectrometric analyses.

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Authors and Affiliations

  1. GI Cancer Research Institute, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, 430030, PR China

    Anyi Liu, Chengxin Yu, Cheng Qiu, Qi Wu, Changsheng Huang, Xiaowei She, Kairui Wan, Lang Liu, Mao Li, Zhihong Wang, Yaqi Chen, Fuqing Hu, Da Song, Kangdi Li, Feng Xu, Senyan Lai, Xuelai Luo, Junbo Hu & Guihua Wang

  2. Department of Immunology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, PR China

    Xun Li

  3. Department of Thyroid and Breast Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, 430030, PR China

    Yaqi Chen

  4. Department of Emergency and Trauma Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, 430030, PR China

    Da Song

  5. Protein Chemistry and Proteomics Facility, Tsinghua University Technology Center for Protein Research, Beijing, 100084, PR China

    Chongchong Zhao & Haiteng Deng

  6. Department of General Surgery, First Affiliated Hospital, School of Medicine, Shihezi University, Shihezi, 832000, PR China

    Xuling Sun

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Contributions

GW and JH designed the research. AL performed the majority of the experiments. QW, YC, CH and XL helped analyze the data. DS and XWS contributed to the bioinformatics analysis. ML and ZW collected clinical samples. CQ and CY performed most of the phenotype experiments. KW, LL and KL assisted in animal experiments. CZ and HD did the mass spectrometry detection and analysis. XLS, SL, XLL and FX provided advice for our study. All authors approved this study.

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Correspondence to Junbo Hu or Guihua Wang.

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Liu, A., Yu, C., Qiu, C. et al. PRMT5 methylating SMAD4 activates TGF-β signaling and promotes colorectal cancer metastasis. Oncogene 42, 1572–1584 (2023). https://doi.org/10.1038/s41388-023-02674-x

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